Summary of Study ST003981
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002492. The data can be accessed directly via it's Project DOI: 10.21228/M8HV6F This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST003981 |
| Study Title | Mitochondrial pyruvate metabolism is disturbed in sepsis |
| Study Summary | To explore changes in mitochondrial pyruvate metabolism in sepsis as a cause for sepsis-associated hyperlactatemia, we performed a 13C pyruvate tracer experiment on isolated liver mitochondria from sham (control) and cecal ligation and puncture (CLP, sepsis) mice. This showed (1) reduced 13C incorporation in all tricarboxylic acid (TCA) metabolites of septic mitochondria proving that mitochondrial pyruvate processing is impaired in sepsis, (2) the mitochondrial pyruvate carrier remains functional as a same amount of labeled pyruvate is measured in both sham and CLP mitochondria, (3) TCA cycle and complex I mediated respiration remain functional in sepsis as the total abundance in TCA metabolites remains unaltered between sham and CLP mitochondria, and (4) mitochondrial pyruvate metabolism is due to impaired pyruvate dehydrogenase activity as a reduced amount of acetyl-CoA was detected in septic mitochondria. |
| Institute | Ghent University |
| Last Name | Libert |
| First Name | Claude |
| Address | Technologiepark-Zwijnaarde 71, 9052 Ghent, Belgium |
| claude.libert@irc.vib-ugent.be | |
| Phone | 09/331.37.00 |
| Submit Date | 2025-06-06 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-08-14 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002492 |
| Project DOI: | doi: 10.21228/M8HV6F |
| Project Title: | Unraveling mitochondrial pyruvate dysfunction to mitigate hyperlactatemia and lethality in sepsis |
| Project Summary: | Sepsis, killing 11 million people yearly, is associated with increased lactate production - a metabolite mechanistically linked to mortality - complicating glucose administration in sepsis. To understand the mechanism behind hyperlactatemia, we applied the cecal ligation and puncture (CLP) model and studied all pyruvate processing routes in liver mitochondria during acute sepsis. Our data suggest that mitochondrial pyruvate-driven respiration is nearly nonexistent in sepsis, not due to insufficient pyruvate uptake or carboxylation but due to a dysfunctional pyruvate dehydrogenase complex (PDC). Septic mitochondria compensate by glutamate-mediated TCA anaplerosis, simultaneously converting some pyruvate into alanine via enhanced mitochondrial glutamic pyruvate transaminase (GPT2) activity. Notably, PDC dysfunction is not caused by PDC inactivation per se but by a shortage of its cofactor, thiamine pyrophosphate (TPP). TPP supplementation restores pyruvate oxidation both ex vivo and in vivo and protects mice from sepsis. TPP also allows safe glucose administration in mice, leading to a novel, robust TPP-plus-glucose therapy. |
| Institute: | Ghent University |
| Department: | Inflammation research center - VIB |
| Laboratory: | Mouse Genetics in Inflammation |
| Last Name: | Libert |
| First Name: | Claude |
| Address: | Technologiepark-Zwijnaarde 71, 9052 Ghent, Belgium |
| Email: | claude.libert@irc.vib-ugent.be |
| Phone: | 09/331.37.00 |
Subject:
| Subject ID: | SU004118 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Treatment condition | Tracer condition |
|---|---|---|---|---|
| SA454333 | MCF001496_LN13 | Liver mitochondria | CLP | 13C pyruvate |
| SA454334 | MCF001496_LN17 | Liver mitochondria | CLP | 13C pyruvate |
| SA454335 | MCF001496_LN01 | Liver mitochondria | CLP | 13C pyruvate |
| SA454336 | MCF001496_LN03 | Liver mitochondria | CLP | 13C pyruvate |
| SA454337 | MCF001496_LN21 | Liver mitochondria | CLP | Buffer |
| SA454338 | MCF001496_LN05 | Liver mitochondria | Sham | 13C pyruvate |
| SA454339 | MCF001496_LN15 | Liver mitochondria | Sham | 13C pyruvate |
| SA454340 | MCF001496_LN07 | Liver mitochondria | Sham | 13C pyruvate |
| SA454341 | MCF001496_LN09 | Liver mitochondria | Sham | 13C pyruvate |
| SA454342 | MCF001496_LN11 | Liver mitochondria | Sham | 13C pyruvate |
| SA454343 | MCF001496_LN19 | Liver mitochondria | Sham | Buffer |
| SA454344 | MCF001496_LN18 | mitochondria supernatant | CLP | 13C pyruvate |
| SA454345 | MCF001496_LN14 | mitochondria supernatant | CLP | 13C pyruvate |
| SA454346 | MCF001496_LN04 | mitochondria supernatant | CLP | 13C pyruvate |
| SA454347 | MCF001496_LN02 | mitochondria supernatant | CLP | 13C pyruvate |
| SA454348 | MCF001496_LN22 | mitochondria supernatant | CLP | Buffer |
| SA454349 | MCF001496_LN06 | mitochondria supernatant | Sham | 13C pyruvate |
| SA454350 | MCF001496_LN16 | mitochondria supernatant | Sham | 13C pyruvate |
| SA454351 | MCF001496_LN10 | mitochondria supernatant | Sham | 13C pyruvate |
| SA454352 | MCF001496_LN08 | mitochondria supernatant | Sham | 13C pyruvate |
| SA454353 | MCF001496_LN12 | mitochondria supernatant | Sham | 13C pyruvate |
| SA454354 | MCF001496_LN20 | mitochondria supernatant | Sham | Buffer |
| Showing results 1 to 22 of 22 |
Collection:
| Collection ID: | CO004111 |
| Collection Summary: | Mice were subjected to the cecal ligation and puncture (CLP) to induce polymicrobial sepsis and the sham (control) procedure. After 24h, liver mitochondria were isolated as described by Frezza et al. (Nature Protocols: 2007;2(2):287-95. doi: 10.1038/nprot.2006.478). Isolated liver mitochondria were spinned down and resuspended in mitochondrial assay solution (MAS, 70 mM sucrose, 220 mM mannitol, 10 mM KH2PO4, 5mM MgCl2, 2 mM HEPES, 1 mM EGTA and 0.2% fatty acid-free BSA) with 13 C pyruvate or only MAS and were incubated for 30 minutes at 37°C in oxygenated conditions. |
| Sample Type: | Mitochondria, Mitochondrial supernatant |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR004127 |
| Treatment Summary: | Mice were subjected to the cecal ligation and puncture (CLP), as described by Rittirsch et al. (Nature Protocols: 2009;4(1):31-6. doi: 10.1038/nprot.2008.214), to induce polymicrobial sepsis and the sham (control) procedure. After 24h, liver mitochondria were isolated. |
Sample Preparation:
| Sampleprep ID: | SP004124 |
| Sampleprep Summary: | Isolated liver mitochondria were spinned down and in mitochondrial assay solution (MAS, 70 mM sucrose, 220 mM mannitol, 10 mM KH2PO4, 5mM MgCl2, 2 mM HEPES, 1 mM EGTA and 0.2% fatty acid-free BSA) with 13C3, 99% sodium pyruvate (10mM; Buchem B.V. CLM-2440-0.5) or only MAS and were incubated for 30 minutes at 37°C in oxygenated conditions. Isolated liver mitochondria and the supernatant of the mitochondria were analyzed by mass spectrometry (LC-MS) to determine total abundance, fractional contribution and 13C-mass isotopologue enrichment of a labeled nutrient (13C3 pyruvate) to the metabolites of interest. A total injection volume of 10 ul/sample was used. |
| Sampleprep Protocol Filename: | Protocol_methods_13C3pyruvate.pdf |
| Processing Storage Conditions: | -80℃ |
Chromatography:
| Chromatography ID: | CH004976 |
| Methods Filename: | Protocol_methods_13C3pyruvate.pdf |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Agilent InfinityLab Poroshell 120 HILIC-Z (150 x 2.1mm, 2.7um) |
| Column Temperature: | 25°C |
| Flow Gradient: | A linear gradient was carried out starting with 90% solvent A and 10% solvent B. From 2 to 12 min the gradient changed to 60% B. The gradient was kept on 60% B for 3 min and followed by a decrease to 10% B. The chromatography was stopped at 25 min |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% acetonitrile |
| Solvent B: | 100% water; 10 mM sodium acetate (pH 9.3) |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN006557 |
| Analysis Type: | MS |
| Analysis Protocol File: | Protocol_methods_13C3pyruvate.pdf |
| Chromatography ID: | CH004976 |
| Num Factors: | 8 |
| Num Metabolites: | 239 |
| Units: | peak intensity |
